The protein kinase C (PKC) isoenzymes expressed by bovine tracheal smooth muscle (BTSM) were identified at the protein and mRNA levels. Western immunoblot analyses reliably identified PKCα, PKCβI and PKCβII. In some experiments immunoreactive bands corresponding to PKCδ, PKCϵ and PKCθ were also labelled, whereas the γ, η and ζ isoforms of PKC were never detected. Reverse transcriptase PCR of RNA extracted from BTSM using oligonucleotide primer pairs designed to recognize unique sequences in the PKC genes for which protein was absent or not reproducibly identified by immunoblotting, amplified cDNA fragments that corresponded to the predicted sizes of PKCδ, PKCϵ and PKCζ, which was confirmed by Southern blotting. Anion-exchange chromatography of the soluble fraction of BTSM following homogenization in Ca2+-free buffer resolved two major peaks of activity. Using ϵ-peptide as the substrate, the first peak of activity was dependent upon Ca2+ and 4β-PDBu (PDBu = phorbol 12,13-dibutyrate), and represented a mixture of PKCs α, βI and βII. In contrast, the second peak of activity, which eluted at much higher ionic strength, also appeared to comprise a combination of conventional PKCs that were arbitrarily denoted PKCα′, PKCβI′ and PKCβII′. However, these novel enzymes were cofactor-independent and did not bind [3H]PDBu, but were equally sensitive to the PKC inhibitor GF 109203X compared with bona fide conventional PKCs, and migrated on SDS/polyacrylamide gels as 81 kDa polypeptides. Taken together, these data suggest that PKCs α′, βI′ and βII′ represent modified, but not proteolysed, forms of their respective native enzymes that retain antibody immunoreactivity and sensitivity to PKC inhibitors, but have lost their sensitivity to Ca2+ and PDBu when ϵ-peptide is used as the substrate.
Conference Abstract| January 01 1985 The Influence of Epithelium on Airway Smooth Muscle Contraction F.M. Cuss; F.M. Cuss 1Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London W12 OHS Search for other works by this author on: This Site PubMed Google Scholar J.D. Palmer; J.D. Palmer 1Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London W12 OHS Search for other works by this author on: This Site PubMed Google Scholar P.J. Barnes P.J. Barnes 1Department of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London W12 OHS Search for other works by this author on: This Site PubMed Google Scholar Clin Sci (Lond) (1985) 68 (s11): 59P. https://doi.org/10.1042/cs068059P Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation F.M. Cuss, J.D. Palmer, P.J. Barnes; The Influence of Epithelium on Airway Smooth Muscle Contraction. Clin Sci (Lond) 1 January 1985; 68 (s11): 59P. doi: https://doi.org/10.1042/cs068059P Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsClinical Science Search Advanced Search This content is only available as a PDF. © 1985 The Biochemical Society and the Medical Research Society1985 Article PDF first page preview Close Modal You do not currently have access to this content.
The local post-buckling behaviour of elliptical hollow section (EHS) tubes under compression is analysed in this paper. It is found that EHS tubes with low to moderate aspect ratios can support loads up to their limit loads but are imperfection sensitive (shell-type behaviour), while EHS tubes with moderate to high aspect ratios can carry loads higher than their limits loads (plate-type behaviour) and are imperfection insensitive. For increasing EHS aspect ratio, it is found that the compressive stresses accumulate near the zones of minimum radius of curvature while the zones of maximum radius of curvature experience a relatively low compressive stress level. Thus, it is likely to apply the effective width concept to EHS tubes with moderate to high aspect ratio.